Solution-based approach to study binding to the eIF4E cap-binding site using CD spectroscopy.

Anal Biochem

Discovery Technology, Hoffmann-La Roche Inc., 340 Kingsland Street, Nutley, NJ 07110, USA.

Published: March 2013

The eukaryotic initiation factor 4E (eIF4E) is the key component of the translational initiation complex that recruits mRNA by binding to a unique "cap" structure located at the 5' end of the mRNA. Overexpression of eIF4E has been implicated in the development of cancer, potentially as a result of increasing the cellular levels of proteins involved in processes that include proliferation and regulation of apoptosis. As a result, the cap-binding site of eIF4E has become a target for the development of anti-cancer therapeutics. The structure of eIF4E bound to the cap mimic 7-methyl-GDP revealed that two tryptophans from different loops in eIF4E sandwiched the 7-methylguanine group between them. This interaction gives rise to a strong exciton coupling signal between the two tryptophans that can be visualized by CD spectroscopy. eIF4E is a challenging protein to work with because of a propensity to aggregate under conditions used in biophysical techniques. CD spectroscopy provides a gentle, solution-based approach to study binding to the cap-binding site of eIF4E. Evidence is provided that the exciton coupling signal can be used to both qualitatively and quantitatively analyze the binding of cap analogs to eIF4E.

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.ab.2012.11.020DOI Listing

Publication Analysis

Top Keywords

cap-binding site
12
eif4e
9
solution-based approach
8
approach study
8
study binding
8
site eif4e
8
exciton coupling
8
coupling signal
8
binding
4
binding eif4e
4

Similar Publications

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!